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Background: Colorectal cancer (CRC) has not only seriously affected people's lives, but also burdened the government healthcare system. Long non-coding RNAs (lncRNA) have attracted more and more attention in the cancer study field. Methods: Experiments were completed in the Medical Research and Innovation Center of Shanghai Pudong Hospital, China from 2019 to 2020. Cell cycle was detected by western blot analyzing and flow cytometry. Apoptosis analysis were determined using flow cytometry or western blot analysis. LncRNA CKMT2-AS1 was knocked down by shRNA transfection. Results: We found CKMT2-AS1 was the most significant=0.0105 for SW480 and P=0.0071 for HCT116) difference lncRNA between colorectal cancer treated with autophagy inducer and colorectal cancer without any treatment. Effective shRNA-CKMT2-AS1 was also designed. Following, we found the treatment of autophagy inducer and autophagy inducer + shRNA-NC were able to suppress the proliferation of both SW480 and HCT116 cells. In addition, the treatment of autophagy inducer + shRNA-CKMT2-AS1 significantly reduced the apoptosis of SW480 and HCT116 cells induced by autophagy. Furthermore, we found the phosphorylation of mTOR, AKT was enhanced in SW480, and HCT116 cells treated with autophagy inducer + shRNA-CKMT2-AS1 compared to the cells treated with autophagy inducer of autophagy inducer + shRNA-NC. Conclusion: Enhancing the expression of CKMT2-AS1 will become a promising strategy to prevent the progress of colorectal cancer.
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Neoplasias Colorretais/metabolismo , Técnicas de Silenciamento de Genes , Glicólise , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Metiltransferases/metabolismo , Proteínas de Neoplasias/metabolismo , Neoplasias Colorretais/genética , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Metiltransferases/genética , Proteínas de Neoplasias/genéticaRESUMO
Increasing evidence has indicated a close association between immune infiltration in cancer and clinical outcomes. However, related research in thyroid cancer is still deficient. Our research comprehensively investigated the immune infiltration of thyroid cancer. Data derived from TCGA and GEO databases were analyzed by the CIBERSORT, ESTIMATE, and EPIC algorithms. The CIBERSORT algorithm calculates the proportions of 22 types of immune cells. ESTIMATE algorithm calculates a stromal score to represent all stromal cells in cancer. The EPIC algorithm calculates the proportions of cancer-associated fibroblasts (CAFs) and endothelial cells (ECs), which are the main components of stromal cells. We analyzed the correlation of immune infiltration with clinical characteristics and outcomes of patients. We determined that the infiltration of CD8+ T cells improved the survival of thyroid cancer patients. Overexpression of immune checkpoints was closely related to the development of thyroid cancer. In general, stromal cells were associated with the progression of thyroid cancer. Interestingly, CAFs and ECs had opposite roles in this process. In addition, the BRAFV600E mutation was related to the upregulation of immune checkpoints and CAFs and the downregulation of CD8+ T cells and ECs. Finally, we constructed an immune risk score model to predict the prognosis and development of thyroid cancer. Our research demonstrated a comprehensive panorama of immune infiltration in thyroid cancer, which may provide potential value for immunotherapy.
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Câncer Papilífero da Tireoide/imunologia , Adulto , Feminino , Humanos , Masculino , Fatores de Risco , Transfecção , Microambiente TumoralRESUMO
Obesity contributes to the incidence of various tumors, including colon cancer. However, the impact of obesity on patients' survival and related mechanisms remains unclear. Multi-omics data of 227 cases of colon cancer patients combined with clinical characteristics data were acquired from The Cancer Genome Atlas (TCGA) database. We confirmed obesity as an independent prognostic factor for improved overall survival of colon cancer patients. We demonstrated that hypoxia pathways were repressed in obese patients by regulating miR-210. Immune checkpoints PD-1 and LAG3 were also downregulated in obese patients, which indicated enhanced immune surveillance. The frequency of PIK3CA and KRAS mutations was decreased in obese patients. The sites and types of TP53 mutation were alternated in obesity patients. In conclusion, our research demonstrated the potential mechanisms of prolonged survival in colon cancer patients combined with obesity, which may provide potential value for improving the prognosis of colon cancer.
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Neoplasias do Colo/genética , Obesidade/genética , Adulto , Antígenos CD/genética , Classe I de Fosfatidilinositol 3-Quinases/genética , Neoplasias do Colo/diagnóstico , Neoplasias do Colo/mortalidade , Neoplasias do Colo/patologia , Bases de Dados Genéticas , Feminino , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , Células HCT116 , Humanos , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Mutação , Obesidade/diagnóstico , Obesidade/mortalidade , Prognóstico , Receptor de Morte Celular Programada 1/genética , Fatores de Proteção , Mapas de Interação de Proteínas , Proteínas Proto-Oncogênicas p21(ras)/genética , Medição de Risco , Fatores de Risco , Transdução de Sinais , Hipóxia Tumoral , Microambiente Tumoral , Proteína Supressora de Tumor p53/genética , Proteína do Gene 3 de Ativação de LinfócitosRESUMO
As shown in our previous study, SIRT6 promotes an aggressive phenotype and the epithelial-mesenchymal transition (EMT) in papillary thyroid cancer (PTC). In this study, we focused on the regulatory axis including SIRT6, autophagy, and the Warburg effect. We innovatively confirmed that SIRT6 overexpression depleted histone H3 lysine 56 acetylation (H3K56ac) of the negative regulator of reactive oxygen species (NRROS) in vitro, thus increasing reactive oxygen species (ROS) production. The accumulated ROS then activated endoplasmic reticulum stress (ER stress) and subsequently induced autophagy. Furthermore, SIRT6 overexpression inhibited glucose transporter 1 (GLUT1) via autophagy-mediated degradation, ultimately suppressing the Warburg effect. Treatment with the ROS scavenger N-acetyl-L-cysteine (NAC, 5 mM) or the autophagy inhibitor chloroquine (CQ) both rescued the inhibition of the Warburg effect. Additionally, a higher concentration of NAC (15 mM) further inhibited the Warburg effect. These concentration-dependent bilateral effects of NAC on this process were confirmed to be due to the regulation of the AMPK signaling pathway. Finally, we further examined this mechanism in vivo by establishing subcutaneous xenografts in nude mice and analyzed the tumors using 18F radio-labeled fluorodeoxyglucose (18F-FDG) PET/CT. In conclusion, we identified a SIRT6-ROS-ER stress-autophagy-GLUT1-Warburg effect axis in PTC, which may provide a new therapeutic target. In addition, NAC (low concentration) and CQ, previously considered to be tumor inhibitors, were shown to promote tumorigenesis in PTC with high SIRT6 expression by inducing the Warburg effect.
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BACKGROUND: N6-Methyladenosine (m6A) modification has been implicated in multiple processes for colon cancer development. IGF2BP3 was a newly reported m6A reader, whereas its role in colon cancer remains unclear. METHODS: The expression of m6A associated enzymes and total m6A level were measured by Western Blotting analysis and m6A RNA Methylation Quantification Kit respectively. Cell cycle was analyzed by flowcytometry. The interaction of IGF2BP3 and related targets was analyzed by RNA immunoprecipitation (RIP) and m6A RNA immunoprecipitation (MeRIP) assays. RESULTS: We investigated all m6A regulated enzymes in colon cancer and found only the overexpression of IGF2BP3 was associated with cancer progression and survival based on The Cancer Genome Atlas (TCGA) databases. Additionally, we also demonstrated IGF2BP3 was associated with DNA replication in the cell cycle. Knockdown of IGF2BP3 significantly repressed percentage of S phase of cell cycle as well as cell proliferation. Further research demonstrated IGF2BP3 bound to the mRNA of Cyclin D1 (CCND1, checkpoint of G1/S phase of cell cycle) and reduced its mRNA stability via reading m6A modification in the CDS region. Overexpression of Cyclin D1 in IGF2BP3 down-regulated cells completely rescued the inhibited percentage of S phase in cell cycle as well as cell proliferation. Additionally, we also demonstrated a similar role of IGF2BP3 at VEGF. IGF2BP3 bound to the mRNA of VEGF and reads m6A modification, thus regulated both expression and stability of VEGF mRNA. Knockdown of IGF2BP3 repressed angiogenesis in colon cancer via regulating VEGF. CONCLUSION: Knockdown of IGF2BP3 repressed DNA replication in the S phase of cell cycle and angiogenesis via reading m6A modification of CCND1 and VEGF respectively. IGF2BP3 was a possible prognosis marker and potential therapeutic target of colon cancer.
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Neoplasias do Colo/genética , Ciclina D1/genética , Neovascularização Patológica/genética , Proteínas de Ligação a RNA/genética , Fator A de Crescimento do Endotélio Vascular/genética , Adenosina/análogos & derivados , Adenosina/genética , Animais , Ciclo Celular/genética , Proliferação de Células/genética , Neoplasias do Colo/patologia , Replicação do DNA/genética , Feminino , Células HCT116 , Xenoenxertos , Humanos , Masculino , Metilação , Camundongos , Neovascularização Patológica/patologia , Processamento Pós-Transcricional do RNA/genética , RNA Mensageiro/genéticaRESUMO
OBJECTIVE: Increasing evidence has indicated an association between immune infiltration in colon cancer and clinical outcomes. The aim of this research is to comprehensively investigate the effect of 22 tumor-infiltrating immune cells (TIICs) on the prognosis of colon cancer patients. METHODS: In our research, CIBERSORT algorithm was used to calculate the proportion of 22 TIICs in 369 colon cancer cases and 39 normal cases from the TCGA cohort. Cox regression analysis was used to analyze the effect of 22 TIICs on the prognosis of colon cancer. Immune risk scoring model was constructed based on the statistical correlation between TIICs subpopulation and survival. Meanwhile, multivariate Cox regression analysis was utilized to investigate whether the immune risk score model was an independent factor for predicting the prognosis of colon cancer. Nomogram was constructed to comprehensively predict the survival rate of colon cancer. P< 0.05 was considered to be statistically significant. RESULTS: The results of the difference analysis showed that except for 12 TIICs, the remaining immune cells exhibited no differential infiltration between normal and colon cancer tissues (p<0. 05). Univariate Cox regression analysis revealed 5 immune cells statistically correlated with colon cancer-related survival risk, including B cells naive, B cells memory, monocytes, macrophages M0, macrophages M1 (P<0.05). In addition, a four-cell based immune risk scoring model was constructed through LASSO Cox regression analysis. KM curve indicated that patients in highrisk were associated with poor outcomes (p<0.001). ROC curve indicated that the immune risk score model was reliable in predicting survival risk (AUC=0.848). Our model showed satisfying AUC and survival correlation in the validation dataset (3-year over survival (OS) AUC=0.941, 5-year OS AUC=0.865, P=0.022). Furthermore, multivariate Cox regression analysis confirmed that the immune risk score model was an independent factor for predicting the prognosis of colon cancer (hazard ratio (HR) =5.017, 95% confidence interval (CI) =2.336-10.777; P<0.001). Ultimately, a nomogram was established to comprehensively predict the survival of colon cancer patients with the results of multivariate Cox regression analysis. CONCLUSION: Collectively, tumor-infiltrating immune cells played an essential role in the prognosis of colon cancer. Furthermore, immune risk score was an independent predictive factor of colon cancer, indicating a poor survival.
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Reactive oxygen species (ROS) is a kind of single electron reduction product of oxygen in vivo. Because they contain unpaired electrons, ROS has high chemical reactivity. Various researches of ROS are focused on DNA damage, cell apoptosis, oxidative stress. Actually, ROS is also closed related to immune regulation. Based on this, we review the research in immune regulated ROS production, regulation of ROS on inflammatory factors and inflammatory response, antigen presentation and macrophage polarization for better understanding of its role.
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Imunomodulação , Espécies Reativas de Oxigênio/metabolismo , Animais , Antioxidantes/metabolismo , Apoptose/imunologia , Biomarcadores , Suscetibilidade a Doenças , Humanos , Macrófagos/imunologia , Macrófagos/metabolismo , Oxirredução , Estresse Oxidativo , Transdução de SinaisRESUMO
Autophagy plays a key role in colorectal cancer (CRC) development and reduces the sensitivity of CRC cells to treatment. The present study reported a novel tumorsuppressive role for autophagy, which was demonstrated to be regulated through the novel oncogene neurotrophin4 (NTF4). NTF4 was significantly overexpressed in tumor tissue compared with nontumor mucosa, and the upregulation of NTF4 in CRC was associated with poor overall survival and advanced TNM stage. The genetic knockdown of NTF4 using short hairpin RNA in CRC cells prevented epithelialtomesenchymal transition and activated autophagy; this was regulated through the interaction between autophagyassociated gene 5 (Atg5) and the mitogenactivated protein kinase pathway. In addition, the knockdown of NTF4 inhibited cell invasion, migration, proliferation and colony formation, and promoted cell cycle arrest. Treatment of the cells with the autophagy inhibitor chloroquine (CQ) rescued these functions and promoted cell invasion, migration, proliferation and colony formation. Finally, the knockdown of NTF4 inhibited the growth of subcutaneous xenografts in Balb/cnu mice. In conclusion, these findings suggested that NTF4 may be a diagnostic marker associated with the overall survival and progression of patients with CRC. NTF4 was found to promote tumorigenesis and CRC development through autophagy regulation.
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Neoplasias Colorretais/patologia , Fatores de Crescimento Neural/genética , Fatores de Crescimento Neural/metabolismo , Regulação para Cima , Animais , Autofagia , Movimento Celular , Proliferação de Células , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Transição Epitelial-Mesenquimal , Feminino , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Células HCT116 , Células HT29 , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Estadiamento de Neoplasias , Transplante de Neoplasias , Análise de SobrevidaRESUMO
Interleukin (IL)1ß is a member of the IL1 family of proteins. IL1 receptor antagonist (IL1RA) is an agent that binds to the IL1 receptor, preventing IL1 from transmitting signals to cells. The present study aimed to identify the role of the IL1ß/1RA axis in epithelialmesenchymal transition (EMT), cell invasion, migration, proliferation and clone formation in colorectal cancer (CRC) and to determine its underlying mechanisms of action. Significantly increased expression of both IL1ß and IL1RA was identified in CRC patient data uploaded in The Cancer Genome Atlas database, and in tumor tissues when compared with matched control tissue. High expression of IL1ß was associated with an increased rate of overall survival and recurrencefree survival. Further research revealed that the IL1ß gene was coexpressed with the IL1RA gene in tumors of CRC patients. It was additionally determined that recombinant human (rh)IL1ß suppressed autophagy as well as EMT in HCT116 cells compared with controltreated cells, whereas rhIL1RA exhibited the opposite effect. In addition, autophagy activator rapamycin (RAPA) rescued the inhibition of EMT in rhIL1ßtreated HCT116 cells. Moreover, rhIL1ß inhibited cell invasion, migration, proliferation and colonyformation ability, when compared with a control treatment. Compared with a control treatment rhIL1RA promoted cell invasion, migration, proliferation, but had no effect on clone formation ability. Furthermore, both rhIL1RA and RAPA rescued inhibition of cell invasion, migration and clone formation ability in rhIL1ßtreated HCT116 cells. RAPA, but not rhIL1RA, rescue inhibited proliferation in rhIL1ßtreated HCT116 cells compared with controls. In addition, it was confirmed that rhIL1ß inhibited the growth of subcutaneous xenografts in nude mice, when compared with control treatments. These results indicated that upregulation of the IL1ß/1RA axis in CRC regulated EMT, cell invasion and migration, proliferation and clone formation via autophagy.
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Proliferação de Células/genética , Neoplasias Colorretais/genética , Proteína Antagonista do Receptor de Interleucina 1/genética , Interleucina-1beta/genética , Idoso , Animais , Apoptose/genética , Autofagia/genética , Movimento Celular/genética , Neoplasias Colorretais/patologia , Transição Epitelial-Mesenquimal/genética , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células HCT116 , Xenoenxertos , Humanos , Masculino , Camundongos , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologiaRESUMO
This study aims to explore the ability of magnetic resonance imaging (MRI) in mucin 1 (MUC1) modified superparamagnetic iron oxide nanoparticle (SPION) targeting human pancreatic cancer (PC). The MUC1 target-directed probe was prepared through MUC1 conjugated to SPION using the chemical method to assess its physiochemical characteristics, including hydration diameter, surface charge, and magnetic resonance signal. The cytotoxicity of MUC1-USPION was verified by MTS assay. BxPC-3 was cultured with MUC1-USPION and SPION in different concentrations. The combined condition of the targeted probes and cells were observed through Prussian blue staining. The nude mice model of pancreatic cancer was established to investigate the application of the probe. MRI was performed to determine the intensity of the signal of the transplanted tumor, while immunohistochemistry and Western blot analysis were performed to detect the expression of MUC1 after taking the transplanted tumor specimen. The particle size of the prepared molecular probe was 63.5 ± 3.2 nm, and the surface charge was 10.2 mV. Furthermore, the probe solution could significantly reduce the MRI at T2 , and the magnetic resonance transverse relaxation rate (ΔR2 ) has a linear relationship with the concentration of iron in the solution. The cell viability of MUC1-USPION in different concentrations revealed no statistical difference, according to the MTS assay. In vitro, the MRI demonstrated decreased T2WI signal intensity in both groups, especially the targeting group. In vivo, MUC1 could selectively accumulate in the nude mice model, and significantly reduce the T2 signal strength. In subsequent experiments, the expression of MUC1 was high in pancreatic cancer tissues, but low in normal pancreatic tissues, as determined by immunohistochemistry and Western blot analysis. The prepared samples can be combined with pancreatic cancer tissue specificity by in vivo imaging, providing reliable early in vivo imaging data for disease diagnosis.
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Aptâmeros de Nucleotídeos/química , Imageamento por Ressonância Magnética/métodos , Nanopartículas de Magnetita/química , Mucina-1/química , Neoplasias Pancreáticas/metabolismo , Animais , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/química , Anticorpos Monoclonais/metabolismo , Aptâmeros de Nucleotídeos/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Meios de Contraste/administração & dosagem , Meios de Contraste/química , Meios de Contraste/metabolismo , Humanos , Imuno-Histoquímica , Nanopartículas de Magnetita/administração & dosagem , Camundongos Nus , Mucina-1/genética , Mucina-1/metabolismo , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/terapia , Tamanho da Partícula , Ensaios Antitumorais Modelo de Xenoenxerto/métodosRESUMO
Interleukin17F (IL17F) is a member of the IL17 family of proteins. Previous studies concerning IL17F have mainly focused on its proinflammatory responses, whereas the present study explored its role as an oncogene in colorectal cancer (CRC). The present study investigated the expression of IL17F in 69 patients with CRC. IL17F was significantly overexpressed in tumor mucosa compared with that in the paired nontumor mucosa. Furthermore, several studies from Gene Expression Omnibus (GEO) databases were analyzed to assess the association between IL17F and overall survival and relapsefree survival time. Recombinant human IL17F protein (rhIL17F) and antiIL17F antibody were used to study the effect of IL17F on the CRC cell line HCT116 in vitro. According to the results of Transwell and wound healing assays, rhIL17F promoted HCT116 cell migration and invasion which was mediated by inducing epithelialmesenchymal transition (EMT), whereas antiIL17F inhibited EMT.
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Biomarcadores Tumorais/metabolismo , Movimento Celular , Neoplasias Colorretais/patologia , Transição Epitelial-Mesenquimal , Regulação Neoplásica da Expressão Gênica , Interleucina-17/metabolismo , Recidiva Local de Neoplasia/patologia , Idoso , Idoso de 80 Anos ou mais , Apoptose , Proliferação de Células , Neoplasias Colorretais/metabolismo , Feminino , Seguimentos , Humanos , Masculino , Invasividade Neoplásica , Recidiva Local de Neoplasia/metabolismo , Prognóstico , Taxa de Sobrevida , Células Tumorais CultivadasRESUMO
Purpose: This study aimed to find risk factors for colon cancer progression with bioinformatics methods, and validated by clinical patients. Methods: Differentially expressed genes (DEGs) between colon cancer tissues and normal colon tissues were extracted from The Cancer Genome Atlas (TCGA) database using R software, amounted to 8,051. DEGs between pathologic stage I+II and stage III+IV amounted to 373, and were compared with DEGs of cancer/normal analyzed above to get the intersection of both. Ninety-six intersected DEGs were identified and defined as progressive DEGs of colon cancer. Then these 96 progressive DEGs were studied by Gene ontology and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis using the DAVID database and visualizing by R software. A protein-protein interaction (PPI) network and functional modules were established using the STRING database. Further, an overall survival (OS) curve was drawn via the GEPIA website based on the CGA database and six progressive DEGs were found to be involved with OS of colon cancer patients. The Linkedomics website was used for detailed analysis of specific subsets of TNM. Results: Pregnancy specific glycoprotein (PSG), vitamin digestion, and absorption were confirmed to promote the progression of colon cancer. Furthermore, NTF4 was found to be associated with both OS and each subset of TNM; therefore, defined as a key risk factor for colon cancer progression. Further analysis of NTF4 expression using clinical data showed it acted as a key risk factor and diagnosis marker for colon cancer progression. Conclusion: NTF4 is a risk factor contributing to colon cancer progression and associated with overall survival.
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Our previous study has demonstrated that knockdown of Grainyhead-like 2(GRHL2) in colorectal cancer (CRC) cells inhibited cell proliferation by targeting ZEB1. This study aimed at researching whether knockdown of GRHL2 promoted CRC progression and metastasis via inducing epithelial-mesenchymal transition (EMT). GRHL2-upregulated SW-620/GRHL2+ and GRHL2-knockdown HCT116/GRHL2-KD, HT29/GRHL2-KD cells and their control cells were generated. The morphological changes after overexpression and knockdown GRHL2 were observed. qRT-PCR, Western blotting, and Immunofluorescence were used to detect EMT markers: E-cadherin, Vimentin, p-catein, ZO-1 and ZEB1 expression. Then, sh-ZEB1 was transfected to GRHL2 knockdown cells to research the relationship between GRHL2 and ZEB1. Transwell and wound healing assays were further performed to detect the impact of GRHL2 on invasion and migration in vitro. CRC cells were injected into mice tail vein to verify the impact of GRHL2 on CRC metastasis. Morphological change of mesenchymal-epithelial transition (MET) could be observed in SW620/GRHL2+ cell. The expression of epithelial markers: E-cadherin, ß-catenin, ZO-1 were up-regulated, while mesenchymal markers: Vimentin was decreased. Meanwhile, opposite EMT morphological change could be observed in HCT116/GRHL2-KD cell, accompanied by reverse change of E-cadherin, ß-catenin, ZO-1, and Vimentin. The expression level of GRHL2 and ZEB1 was found negative in both SW620/GRHL2+ and HCT116/GRHL2-KD cells. Knockdown of ZEB1 by siRNA in HCT116/GRHL2-KD and HT29/GRHL2-KD could upregulate expression of E-cadherin and GRHL2. GRHL2 knockdown also promoted migration, invasion in vitro and CRC metastasis in mice model. In conclusion, GRHL2/ZEB1 axis inhibits CRC progression and metastasis via oppressing EMT.
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Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Proteínas de Ligação a DNA/genética , Transição Epitelial-Mesenquimal/genética , Fatores de Transcrição/genética , Animais , Biomarcadores , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Neoplasias Colorretais/metabolismo , Modelos Animais de Doenças , Progressão da Doença , Receptores ErbB/metabolismo , Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Camundongos , Metástase Neoplásica , Estadiamento de Neoplasias , Transdução de Sinais , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Purpose: Our previous study demonstrated that SIRT6 is upregulated in papillary thyroid cancer (PTC) and enhances tumor aggressiveness. In this study, we further researched its influence in the Warburg effect. Methods: SIRT6-upregulated and downregulated BCPAP cells and negative control BCPAP-NC groups were generated with lentiviral vectors. In these two cell lines, reactive oxygen species (ROS) were detected by dichlorodihydrofluorescein diacetate. Expression of the key Warburg effect genes including GLUT1, HK2, GAPDH, PGK1, ENO1, PKM2 and LDHA was measured by quantitative real-time PCR and western blotting. Glucose uptake, lactate production and the ATP content of cells were detected with assay kits. The ROS scavenger N-acetylcysteine was used for treatment of BCPAP-SIRT6, and the same measurements as described above were detected again. Results: Compared with the BCPAP-NC group, expression of the key Warburg effect genes including Glut1, HK2 and GAPDH and their protein products was upregulated in the BCPAP-SIRT6 group, whereas BCPAP-shSIRT6 showed significant downregulation. Meanwhile, ROS, glucose uptake, lactate production and ATP content of the BCPAP-SIRT6 group were also significantly increased, and BCPAP-shSIRT6 showed significant downregulation. Furthermore, upregulation of key Warburg effect genes and glucose uptake, lactate production and ATP content were all rescued after treatment with ROS scavenger. Conclusion: SIRT6 promoted the Warburg effect of PTC cells via upregulation of ROS. Inhibition of ROS in SIRT6-upregulated cells could rescue activation of the Warburg effect.
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BACKGROUND: In our previous study, we demonstrated that Sirtuin 6 (SIRT6) is upregulated and associated with papillary thyroid cancer (PTC) progression (Qu et al. in Int J Oncol 50(5):1683-92, 2017). This study examined whether SIRT6 promotes epithelial-mesenchymal transition (EMT) of papillary thyroid cancer through hypoxia inducible factor-1α (HIF-1α). METHODS: SIRT6-upregulated TPC-1 and B-CPAP cells were generated by lentivirus. Western blotting, RT-qPCR, immunofluorescence was performed to detect the following EMT associated markers: E-cadherin, Vimentin, Snail, and TWIST. Cell proliferation was detected by CCK8, and cell invasion and migration were detected by transwell and wound healing assays, respectively. HIF-1α expression was further detected by western blotting in both normoxia and hypoxia conditions. A HIF-1α inhibitor was then used to block HIF-1α expression in SIRT6-upregulated PTC cells. The same parameters were then assessed and compared with control HIF-1α cells. RESULTS: E-cadherin was significantly decreased, whereas Vimentin, Snail, and TWIST were increased in SIRT6-upregulated PTC cells. Additionally, SIRT6 promoted the invasion and migration of PTC cells. We found that SIRT6 enhanced HIF-1α stability and synthesis and prolonged the protein half-life. The changes in the EMT associated markers and in the invasion and migration ability were rescued after inhibition of HIF-1α expression. Furthermore, we found that SIRT6 increased PTC resistance to HIF-1α inhibitor-mediated proliferation changes. CONCLUSION: These results confirm that the SIRT6/HIF-1α axis promotes papillary thyroid cancer progression by inducing EMT.
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At present there is no consensus on the treatment of classical Hodgkin's lymphoma (CHL) following relapse. The aim of the present study was to access the class I-selective histone deacetylase (HDAC) inhibitor (HDACI) MGCD0103 on the expression levels of Bcl-2, nuclear factor (NF)-κB and programmed death-ligand 1 (PD-L1) in CHL, to explore the possible therapeutic value of MGCD0103 in combined relative target drugs for patients with CHL. In L1236 and L428 cell lines, apoptosis and cell cycle stage were identified using flow cytometry, and the effects of HDACI on CHL were assessed in terms of Bcl-2, NF-κB and PD-L1 expression levels, which were detected by western blotting and co-focusing experiments. The results demonstrated that MGCD0103 could induce cell apoptosis and cell cycle arrest, down-regulate Bcl-2 and increase NF-κB and PD-L1 expression levels in L1236 and L428 cell lines. MGCD0103 decreases Bcl-2 levels and upregulates PD-L1, which indicates that the combined use of HDACIs and a PD-L1 inhibitor in theory may improve treatment outcomes in patients with CHL. MGCD0103 may also up-regulate NF-κB, which seems to induce resistance towards anti-apoptotic drugs. Clinical trials combining HDACIs with NF-κB and/or PD-L1 inhibitors should be designed to further improve treatment outcomes for patients with CHL.
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Histone deacetylases (HDACs) are involved in multiple physical and pathological processes in classical Hodgkin lymphoma (cHL). The prognostic value of HDACs in cHL patients has not been discussed. The aim of the current study is to investigate the HDAC1, HDAC2, HDAC3, and HDAC11 expressions, and to evaluate the correlation of HDAC1, HDAC2, HDAC3, and HDAC11 expressions with the survival rate in cHL patients. We retrospectively analyzed clinicopathological data of 28 patients who were diagnosed with cHL between August 2002 and March 2010. Immunohistochemistry was used to detect the expression of HDAC1, HDAC2, HDAC3, and HDAC11 in these patients. The results showed that HDAC1, HDAC3, and HDAC11 were expressed at a higher level in Hodgkin Reed-Sternberg cells, whereas HDAC2 was expressed at a lower level in Hodgkin Reed-Sternberg cells. The expression of HDAC2 had a relationship with pathological type (P=0.012). There was also a correlation between the expression of HDAC11 and the erythrocyte sedimentation rate (P=0.054). Other clinicopathological parameters had no significant correlation with the expression of HDAC1, HDAC2, HDAC3, and HDAC11 in terms of survival (P>0.05). The 10-year total survival rate by Cox multivariate analysis, after taking into account all clinical and pathologic factors, showed that bulky disease retained significance (P=0.028). Higher expression of HDAC1 predicted shorter progression-free survival and overall survival (OS) in cHL patients (P<0.05, in both cases), and higher expression of HDAC11 might be correlated with lower OS (P=0.05). The study showed that the expressions of HDAC2 and HDAC11 have a particular relationship with the pathologic subtype. Increased expression of HDAC1 was correlated negatively with progression-free survival and OS, and increased expression of HDAC11 had a borderline relationship with the OS rate in patients with cHL.
Assuntos
Histona Desacetilase 1/biossíntese , Histona Desacetilase 2/biossíntese , Histona Desacetilases/biossíntese , Doença de Hodgkin/enzimologia , Adulto , Biomarcadores Tumorais , Tratamento Farmacológico , Feminino , Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Doença de Hodgkin/patologia , Doença de Hodgkin/terapia , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Intervalo Livre de Progressão , Radioterapia Adjuvante , Estudos RetrospectivosRESUMO
BACKGROUND: Gastric plication (GP) and sleeve gastrectomy (SG) are 2 restrictive bariatric surgeries that are associated with weight loss and health improvement in patients with obesity. However, differences in how these procedures exert their effects have not been systematically evaluated and compared between techniques. OBJECTIVES: To investigate the effectiveness of GP and SG surgeries for obese patients with type 2 diabetes based on evaluation of energy metabolism, hormone metabolism, and gastrointestinal dynamics. SETTING: University medical center. METHODS: Zucker diabetic fatty rats (n = 30) were equally and randomly divided into 3 groups: sham, GP, and SG. Weight, food intake, fasting plasma glucose (FPG), and intraperitoneal glucose tolerance test (IPGTT) were measured in vivo before operation and at 2, 4, and 6 weeks postoperation. Whole-body metabolic parameters including activity, energy expenditure, and respiratory exchange rate (RER) were measured using metabolic cages 3 weeks postoperation. Blood samples were taken 2 weeks before operation and at 2, 4, and 6 weeks postoperation for the purpose of measuring the expression of serum ghrelin and glucagon-like peptide (GLP-1) by enzyme-linked immunosorbent assay. The residual gastric and intestinal propulsive movement were measured at 6 weeks postoperation after all animals were sacrificed. RESULTS: Compared with sham, the GP and SG procedures achieved near equivalent levels of efficacy as far as weight loss, reduced food intake, and decreased FBG and IPGTT in our rodent model. The GP and SG procedures also provided the same effectiveness as far as altering serum ghrelin and GLP-1 hormones. In addition, results showed that the GP and SG procedures can increase metabolic rate by consuming more energy and reducing activity. RERs in GP and SG animals were lower than in sham animals, which indicates that the energy mainly comes from adipose tissue. Moreover, the GP procedure showed lower gastric residual compared to sham, while the SG procedure did not appear to have this affect; the SG procedure resulted in deficiencies in intestinal propulsion function. CONCLUSIONS: The GP and SG procedures have the same effectiveness and can help improve diabetes control by regulating weight, glucose tolerance, and metabolic hormones and augmenting gastrointestinal dynamics. Therefore, these procedures have great potential as therapies for obesity and type 2 diabetes.